Monitoring of gene expression in bacteria during infections using an adaptable set of bioluminescent, fluorescent and colorigenic fusion vectors.

2.50
HDL Handle:
http://hdl.handle.net/10033/202649
Title:
Monitoring of gene expression in bacteria during infections using an adaptable set of bioluminescent, fluorescent and colorigenic fusion vectors.
Authors:
Uliczka, Frank; Pisano, Fabio; Kochut, Annika; Opitz, Wiebke; Herbst, Katharina; Stolz, Tatjana; Dersch, Petra
Abstract:
A family of versatile promoter-probe plasmids for gene expression analysis was developed based on a modular expression plasmid system (pZ). The vectors contain different replicons with exchangeable antibiotic cassettes to allow compatibility and expression analysis on a low-, midi- and high-copy number basis. Suicide vector variants also permit chromosomal integration of the reporter fusion and stable vector derivatives can be used for in vivo or in situ expression studies under non-selective conditions. Transcriptional and translational fusions to the reporter genes gfp(mut3.1), amCyan, dsRed2, luxCDABE, phoA or lacZ can be constructed, and presence of identical multiple cloning sites in the vector system facilitates the interchange of promoters or reporter genes between the plasmids of the series. The promoter of the constitutively expressed gapA gene of Escherichia coli was included to obtain fluorescent and bioluminescent expression constructs. A combination of the plasmids allows simultaneous detection and gene expression analysis in individual bacteria, e.g. in bacterial communities or during mouse infections. To test our vector system, we analyzed and quantified expression of Yersinia pseudotuberculosis virulence genes under laboratory conditions, in association with cells and during the infection process.
Affiliation:
Department of Molecular Infection Biology, Helmholtz Centre for Infection Research, Braunschweig, Lower Saxony, Germany.
Citation:
Monitoring of gene expression in bacteria during infections using an adaptable set of bioluminescent, fluorescent and colorigenic fusion vectors. 2011, 6 (6):e20425 PLoS ONE
Journal:
PloS one
Issue Date:
2011
URI:
http://hdl.handle.net/10033/202649
DOI:
10.1371/journal.pone.0020425
PubMed ID:
21673990
Type:
Article
Language:
en
ISSN:
1932-6203
Appears in Collections:
Publications of Molekulare Infektionsbiologie(MIBI)

Full metadata record

DC FieldValueLanguage
dc.contributor.authorUliczka, Franken
dc.contributor.authorPisano, Fabioen
dc.contributor.authorKochut, Annikaen
dc.contributor.authorOpitz, Wiebkeen
dc.contributor.authorHerbst, Katharinaen
dc.contributor.authorStolz, Tatjanaen
dc.contributor.authorDersch, Petraen
dc.date.accessioned2012-01-12T13:40:14Z-
dc.date.available2012-01-12T13:40:14Z-
dc.date.issued2011-
dc.identifier.citationMonitoring of gene expression in bacteria during infections using an adaptable set of bioluminescent, fluorescent and colorigenic fusion vectors. 2011, 6 (6):e20425 PLoS ONEen
dc.identifier.issn1932-6203-
dc.identifier.pmid21673990-
dc.identifier.doi10.1371/journal.pone.0020425-
dc.identifier.urihttp://hdl.handle.net/10033/202649-
dc.description.abstractA family of versatile promoter-probe plasmids for gene expression analysis was developed based on a modular expression plasmid system (pZ). The vectors contain different replicons with exchangeable antibiotic cassettes to allow compatibility and expression analysis on a low-, midi- and high-copy number basis. Suicide vector variants also permit chromosomal integration of the reporter fusion and stable vector derivatives can be used for in vivo or in situ expression studies under non-selective conditions. Transcriptional and translational fusions to the reporter genes gfp(mut3.1), amCyan, dsRed2, luxCDABE, phoA or lacZ can be constructed, and presence of identical multiple cloning sites in the vector system facilitates the interchange of promoters or reporter genes between the plasmids of the series. The promoter of the constitutively expressed gapA gene of Escherichia coli was included to obtain fluorescent and bioluminescent expression constructs. A combination of the plasmids allows simultaneous detection and gene expression analysis in individual bacteria, e.g. in bacterial communities or during mouse infections. To test our vector system, we analyzed and quantified expression of Yersinia pseudotuberculosis virulence genes under laboratory conditions, in association with cells and during the infection process.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshArtificial Gene Fusionen
dc.subject.meshBacteriaen
dc.subject.meshBacterial Infectionsen
dc.subject.meshBase Sequenceen
dc.subject.meshCell Lineen
dc.subject.meshFemaleen
dc.subject.meshGene Expression Regulation, Bacterialen
dc.subject.meshGenetic Vectorsen
dc.subject.meshLuminescent Proteinsen
dc.subject.meshMiceen
dc.subject.meshPlasmidsen
dc.subject.meshPromoter Regions, Geneticen
dc.subject.meshYersinia pseudotuberculosisen
dc.titleMonitoring of gene expression in bacteria during infections using an adaptable set of bioluminescent, fluorescent and colorigenic fusion vectors.en
dc.typeArticleen
dc.contributor.departmentDepartment of Molecular Infection Biology, Helmholtz Centre for Infection Research, Braunschweig, Lower Saxony, Germany.en
dc.identifier.journalPloS oneen

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